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Crosstalk between hydrogen sulfide and nitric oxide in endothelial cells.

Altaany Z, Yang G, Wang R - J. Cell. Mol. Med. (2013)

Bottom Line: Their interaction at different levels would have a profound impact on angiogenesis.Blockade of NO production by eNOS-specific siRNA or nitro-L-arginine methyl ester (L-NAME) reversed, but eNOS overexpression potentiated, the proliferative effect of H2 S on ECs.Our results suggest that H2 S stimulates the phosphorylation of eNOS through a p38 MAPK and Akt-dependent pathway, thus increasing NO production in ECs and vascular tissues and contributing to H2 S-induced angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Lakehead University, Thunder Bay, Ontario, Canada.

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H2S stimulated the phosphorylation of endothelial NO synthase (eNOS) in endothelial cells (ECs). (A) The effect of NaHS treatment on eNOS phosphorylation. ECs were starved in Dulbecco's modified eagles medium (DMEM) medium free of serum for 24 hrs and treated with different concentrations of NaHS for 30 min. Western blot analysis was conducted using anti-phospho-eNOS and anti-total eNOS antibody, n = 3–4, *P < 0.05 versus control. (B) Time-dependent effect of NaHS treatment on the phosphorylation of eNOS. ECs were treated with NaHS (100 μM) for different periods (0–60 min.). At the end of each time-point, cells were collected and proteins lysates were analysed by Western blot, n = 3–4, *P < 0.05 versus control. (C) The effect of NaHS treatment on eNOS expression level in ECs. The ECs were treated with NaHS (100 μM) for 12–36 hrs, and then cells were collected and proteins were subjected to Western blot analysis. n = 3–4, *P < 0.05 versus control.
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fig02: H2S stimulated the phosphorylation of endothelial NO synthase (eNOS) in endothelial cells (ECs). (A) The effect of NaHS treatment on eNOS phosphorylation. ECs were starved in Dulbecco's modified eagles medium (DMEM) medium free of serum for 24 hrs and treated with different concentrations of NaHS for 30 min. Western blot analysis was conducted using anti-phospho-eNOS and anti-total eNOS antibody, n = 3–4, *P < 0.05 versus control. (B) Time-dependent effect of NaHS treatment on the phosphorylation of eNOS. ECs were treated with NaHS (100 μM) for different periods (0–60 min.). At the end of each time-point, cells were collected and proteins lysates were analysed by Western blot, n = 3–4, *P < 0.05 versus control. (C) The effect of NaHS treatment on eNOS expression level in ECs. The ECs were treated with NaHS (100 μM) for 12–36 hrs, and then cells were collected and proteins were subjected to Western blot analysis. n = 3–4, *P < 0.05 versus control.

Mentions: NaHS (50 and 100 μM) treatment markedly increased the phosphorylation of eNOS in ECs (Fig. 2A). The stimulatory effect of NaHS on eNOS phosphorylation was time dependent, and the increase in phosphorylated eNOS appeared at 10 min., peaked at 30 min., and gradually declined to baseline over the period of 1-hr NaHS exposure (Fig. 2B). NaHS treatment up to 36 hrs had no significant effect on eNOS expression level (Fig. 2C).


Crosstalk between hydrogen sulfide and nitric oxide in endothelial cells.

Altaany Z, Yang G, Wang R - J. Cell. Mol. Med. (2013)

H2S stimulated the phosphorylation of endothelial NO synthase (eNOS) in endothelial cells (ECs). (A) The effect of NaHS treatment on eNOS phosphorylation. ECs were starved in Dulbecco's modified eagles medium (DMEM) medium free of serum for 24 hrs and treated with different concentrations of NaHS for 30 min. Western blot analysis was conducted using anti-phospho-eNOS and anti-total eNOS antibody, n = 3–4, *P < 0.05 versus control. (B) Time-dependent effect of NaHS treatment on the phosphorylation of eNOS. ECs were treated with NaHS (100 μM) for different periods (0–60 min.). At the end of each time-point, cells were collected and proteins lysates were analysed by Western blot, n = 3–4, *P < 0.05 versus control. (C) The effect of NaHS treatment on eNOS expression level in ECs. The ECs were treated with NaHS (100 μM) for 12–36 hrs, and then cells were collected and proteins were subjected to Western blot analysis. n = 3–4, *P < 0.05 versus control.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3822893&req=5

fig02: H2S stimulated the phosphorylation of endothelial NO synthase (eNOS) in endothelial cells (ECs). (A) The effect of NaHS treatment on eNOS phosphorylation. ECs were starved in Dulbecco's modified eagles medium (DMEM) medium free of serum for 24 hrs and treated with different concentrations of NaHS for 30 min. Western blot analysis was conducted using anti-phospho-eNOS and anti-total eNOS antibody, n = 3–4, *P < 0.05 versus control. (B) Time-dependent effect of NaHS treatment on the phosphorylation of eNOS. ECs were treated with NaHS (100 μM) for different periods (0–60 min.). At the end of each time-point, cells were collected and proteins lysates were analysed by Western blot, n = 3–4, *P < 0.05 versus control. (C) The effect of NaHS treatment on eNOS expression level in ECs. The ECs were treated with NaHS (100 μM) for 12–36 hrs, and then cells were collected and proteins were subjected to Western blot analysis. n = 3–4, *P < 0.05 versus control.
Mentions: NaHS (50 and 100 μM) treatment markedly increased the phosphorylation of eNOS in ECs (Fig. 2A). The stimulatory effect of NaHS on eNOS phosphorylation was time dependent, and the increase in phosphorylated eNOS appeared at 10 min., peaked at 30 min., and gradually declined to baseline over the period of 1-hr NaHS exposure (Fig. 2B). NaHS treatment up to 36 hrs had no significant effect on eNOS expression level (Fig. 2C).

Bottom Line: Their interaction at different levels would have a profound impact on angiogenesis.Blockade of NO production by eNOS-specific siRNA or nitro-L-arginine methyl ester (L-NAME) reversed, but eNOS overexpression potentiated, the proliferative effect of H2 S on ECs.Our results suggest that H2 S stimulates the phosphorylation of eNOS through a p38 MAPK and Akt-dependent pathway, thus increasing NO production in ECs and vascular tissues and contributing to H2 S-induced angiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Lakehead University, Thunder Bay, Ontario, Canada.

Show MeSH